1. Field of the Invention
This invention relates to synthetic genes and their expression products. Specifically, this invention relates to a synthetic protease gene and its expression product.
2. Description of the Related Art
The presence of protease protein in purified virion preparation was shown only by immunological techniques. The HIV protease sequence together with the gag and pol sequence of fusion proteins have been expressed from viral DNA in bacteria. Examples of each disclosures include: 1. Henderson, et al., 1988, "Human Retroviruses, Cancer and AIDS: Approaches to Prevention and Therapy", D. Bolognesi Ed. Published by Alan R. Liss Inc., New York, N.Y. pp. 135-147; 2. Dubouck, et al., 1987, [P.N.A.S., 84:8903-8906; and 3. Mous, et al., 1988, J. Virol, 62:1433-1436.
The primary sequences of the HIV protease has been determined by protein analysis and by the nucleotide sequence of the proviral DNA. It was thus determined that the protease is a 99 amino acid long protein encoded by a 297 bp long stretch of the HIV provirus. All previous experiments on the protease gene and on its expression were carried out by utilizing nucleotide sequences cloned out from the cDNA of the provirus. The inventors' work using synthetic DNA proves that the nucleotide sequence of the provirus DNA and also the deduced amino acid sequence are correct.
The complete nucleotide sequence of the HIV-1 proviral DNA was published by Ratner et al., 1985, Nature, 313:277-284. The sequence coding for the protease in the pol open reading frame of HIV was determined by previous analysis and corresponds to nucleotide 1833 to 2129. The N terminus and the C terminal amino-acids are proline and phenylalanine respectively. This sequence coding for the HIV-I 99 aminoacid protease is 297 bp long as follows. ##STR1##
The industry is lacking a synthetic DNA sequence that encodes a specific enzyme or protease which is essential for the completion replication of an infective human immunodeficiency virus (HIV). This DNA sequence is desirable to express this protease by recombinant methodology in bacteria and or in eukaryotic cells, and to produce enough protease for biochemical and physical characterization in order to design and produce potent inhibitors of this enzyme, and thereby to block the production of infective HIV particles.